Preservation of biologically active and other substances



c. H. BARR 2.198.752

PRESERVATION OF BIOLOGIGALLY ACTIVE AND OTHER SUBSTANCES April so,1940.-

Filed July io 3 Sheets-Sheet l l lf |NVENTOR ATTORNEYS c. H. BARR2,198,752

PRESERVATION OF BIOLOGICALLY ACTIVE AND THER SUBSTANCES April 3o,` 194o.

Filed July l0, 1936 .3 Sheets-Sheet 2 m C h .w

AApril 30, 1940. l c, H. BARR PRESERVATION OF BIOLOGICALLY ACTI-VE ANDOTHER SUBSTANCES 5 Sheets-Shee't 3 Filed July l0, 1936 ATTORNEYSPatented Apr. so, 1940 UNITED STATES PRESERVATION 0F' BILOGICALLY ACTIVEAND OTHER SUBSTANCES Courtland li. Barr, Glenolden, Pa., alaignor toSharp a Dohme, Incorporatcd,`lhlladelphia, Pa., a corporation ofMaryland (1929) Application July 10, 1936, Serial No. 90,033

11 Claims.l (Cl. 226-82) The present invention relates to improvementsin the preservation of biologically active and other substances,including such substances as sera, protein solutions, bacterialcultures, phar- 5 maceutical and glandular substances, viruses and otherlabile biological substances; and it relates more particularly toimprovements in methods and apparatus for sealing such substances infinaln Desiccated biologically active substances are very porous, and itisdesirable to package and distribute them under a vacuum, both toprevent any deterioration which might be caused by contact with the air.and to facilitate the dissolving I of the material in water or otheraqueous nuid when restoration is desired. If such material is maintainedunder a vacuum, and water is introduced into the container before thevacuum is broken, the vacuum tends to pull the water n into the poresand interstices of the material, insuring intimate contact of the waterwith the material and preventing the material from being impregnatedwith gas which might tend to prevent the ready access of water to theinterior .wportionsof the .desiccatcd material. Methods of producingsuch desiccated biologically active sub-` mstances, in iinal containers,sealed under a high vacuum in containers provided with all-glass seals,and perforable closures sealed within the n glass have been developed byDr. Reichel of the Mulforr Laboratories. Such containers enable thedistribution and storage of the materials while maintained under a highvacuum, with absolute assurance that the vacuum is' maintained 45 andthat the material is kept free from contamination, and yet permit waterto' be introduced without breaking the vacuum, by breaking the glassseal over the perforable closure, and introducing water, by means of ahollow needle u, through this closure, which serves to hold the vacuumafter the glass seal is broken.

It is an object ofthe present invention to providean improved processand apparatus by which desiccated biologically active and other "Lsubstances may be produced and stored in a container sealed with anall-glass seal and with a periorable stopper, such as a rubber stopper,sealed within the glass; and to provide such a container, having a bodyportion and a single neck, with a perforable stopper sealed within 5 theneck, the material being sealed within the container under the originalvacuum, such that there is absolute assurance that the vacuum ismaintained during storage, etc., and that the material is kept free fromcontamination, air, moisl0 ture, etc., and such that, when it is desiredto restore the material toa liquid state, water or vother liquid may beintroduced without destroying the vacuum.

In accordance with Ythe present invention, a ll suitable vial orcontainer of ordinary soft glass, preferably cylindrical in shape,provided with a relatively. long tubular neck, the inside of which atthe point adjacent lthe top of the vial or container is tapered to nt arubber'stopper. is pron vided. 'Ihe size of the vial may be variedalmost at will, its capacity being determined by the amount of materialwhich itis intendedto contain and being .such as to contain one or moreunit portions of the material. Thus the capacity of the container mayvary from a fraction of a cubic centimeter to 50 or 100 or more cubiccentimeters.

Where the container is intended for the storage and distribution ofdesiccated biologically active substances, which are to be desiccated inthe container, its volume should be somewhat more than twice as great asthe volume of the material intended to be processed therein, in .orderto provide adequate surface for the sublimation or evaporation of waterthereirom... vThus if unconcentrated material is processed in thecontainer, the`u container must have a volume `of about twice the volumeof the nal restored product, whereas if concentrated material, such asman terial which has been concentrated to about onel half volume in asuitable manner, is processed within the container, the container mayhave an amount of desiccated material within it which on restoration toits normal liquid condition about illls the container. 'I'he .neck ofthe vial.'

evenwhere tapered, must be of suilicient size 25 as a result, thematerial is sealedw'ithin 'the c'onmosphere or surroundings.

the ow of heat into the material from the at- In order to maintain thesublimation or vaporizat'ion at `a sufciently rapid rate to prevent themelting of the material, it is necessary to provide an adequately largepassageway for the vapors, and to avoid the use of vapor passages' oftoo small a lumen, or with too many constrictions.

Into lhe'container so provided is introduced the proper amount ofbiologically active sub- I stance to'be treated, and this substance isthen frozen, preferably while the container is in a .more or lesshorizontal position to insure a' maxi- `mum exposed surface forsublimation, and by ex" posure to a refrigerant maintained at a very lowtemperaturee, g. .-70" C. The container is then attached to a vacuummanifold and subjected tothe action of a, high vacuum to remove thecation has proceeded to the desired extent, a

waterzfrom the frozen material. After the desicl' rubber stopper isintroduced into the neck of the vial anjd forced down to form a tightjoint in the tapered portion of the neck of vial or container. This isdone without breaking the vacuum, and

tainer under a high vacuum with a perforable closure.

The glass neck of the vial, which extends considerably above the rubberstopper, is 'then flame-sealed,` either while the vial is still at'-tached to the vacuum manifold, or after removing the^vial vfrom themanifold and drawing a portion of the neck to a. capillary tube,re-evacuating` thel neck and flame-sealing' the neck at the point whereit has been drawn` to a fine tube. It is advantageous to draw a portionof the neck at the point of sealing to a fine tube before sealing asthis simplifies the sealing operation, and insures the production of avacuum tight seal, without risk of the glass'puncturing, as frequentlyoccurs when tubes of Arelatively large diameter, made from soft glass,are flame- Vsealed under a vacuum. After the sealing, orbefore, the tubeor neck of the vial is etched or scratched a little below `thel top ofthe rubber stopper, sov that it may be readily broken" off at this:`point. The desiccated material so produced maintained as a formed porousmass, having the shape and volume of the frozen material from which ismaintained within the container, during 'storage by means of the glassseal, and. just prior to restoration by means of the rubber stopper,facilitates the introduction of water and insures the complete andspeedy dissolving of the material in the water. l

lT o carry out this process, and produce the lfinal container containingthe treated material, the invention provides apparatus of suitableconstruction. In general, the apparatus comprises an attachment orapparatus suitably connected to a vacuum manifold, and provided withmeans for attaching containers, containing frozen material to bedesiccated,.to it. The attachment also provides a passageway ofrelatively large duced;

'onto the threaded sleeve.

to maintain thematerial ina frozen state despite diameter to. permit thefree flow of vapors from the' container to @the vacuum manifold. Theattachment also includes means by which rubber Stoppers may be readilyintroduced into the neck of the vial after the desiccation is complete,and means by'which the Stoppers may be forced into proper position toinsure the production of a tight seal and the maintenance of a highvacuum within the container even though the neckof the container is openand exposed to atmospheric pressure for relatively short periods oftime, as in flame-sealing or in restoring the desiccated material ,to aliquid state.

The invention will be Afurther illustrated and explained inconnectionwith the drawings, which villustrate an attachment or apparatus whichembodies the invention and which illustrate a container suitable f formarketing and distribution containing a desiccated biologically activesubstance which embodies the invention, and which illustrate the meansby which water may be introduced into such container to restore thematerial to a liquid state without breaking the vacuum lmaintainedwithin the container.

In the drawings, .l Fig, 1 is a sectional view of the attachment orapparatus shown inthe position used during the desiccation of thelfrozen material;

Fig.- 2 is a sectional. view of a portion of the apparatus or attachmentshowing the means byA which a stopper is introducedinto the neck of thevial or container;

Fig. 3 is a sectional view of a portion of the apparatus or attachmentshowing the means by which the rubber stopper is forced' into place toform a tight seal;

Figs. 4 and 5 are views showing the final container before and after theglass seal is made;

Figs.l 6, 7 and 8 are views showing the conl tainer after the glass sealis broken 01T and when the container is ready for the introduction ofwater and means by which water may be intro- Fig. 9 is a view inperspective, showing the attachment fastenedto a vacuum manifold;

Fig. 10 is a `view of a .portion of a suitable vacuum manifold, showinga suitable arrangement ofoutlets and means for'fastening the attachmentsthereto; and

Fig.`11 is a view showing the final container with a portion of the neckabove the stopper drawn to a ne tube; and l Fig. ,12 isaview showing thesealing of the neck above the stopper while maintaining a vacuum in theneck.

In Fig. 1, l represents a vial or container of suitable size intendedfor the marketing or distribution of av desiccated serum or otherbiologically4 active substance, containing a frozen or desiccatedbiologically active substance la, provided with a relative long neck 2of suitable diameter and with a tapered portion 3 at the top of the vialor container. This vial or container isv tion of the rubber tube is thenturned back on itself as shown at 1, and a brass cap 9 screwed l Theportion of the rubber tube under the cap is thus of double thickness,and the cap limits the expansion of this portion of the tube. 4When theneck of a container 'I'he metal tube 4 is provided with a side outlet Iextending upwardly at an angle.

Within tube 4 is located a hollow plunger II, which may be provided witha number of openings I Ia, through which the vapors or gases from thecontainer I flow to the vacuum manifold. 'I'his hollow plunger isrigidly connected to metal tube I2, as at I2a. The tube I2 is alsoconnected to tube 4 by means of an extensible tube I3, which consists ofan inner spiral or metal spring 4, and an outer or surrounding rubbertube I5, and which is readily lengthened or shortened. A stop I 4a,which may have a number of holes I4b, is provided at the bottom of thetube I 2to confine the spring I4.

'I'he plunger I I is of sulcient length to normally block the openingbetween the side outlet I8 and sleeve 4. The metal spring or spiral isprovided to prevent the rubber tube, which is rel-` atively long andmust be extensible, from collapsing under the influence of the vacuum.The connections between tubes 4 and I2 and tube I3 are, of course,vacuum-tight. Tube I2 is connected to thev vacuum pump or vacuummanifold by metal tube I6 and the rubber tube I1, which is made ofrelatively heavy rubber, and which may be clamped to permit the sealingoff of the lower part of the apparatus from the vacuum pump or manifoldas when disconnecting the vial or container from the attachment, etc.Attached to the side outlet I8 by means of the rubber connection I 8, isa metal tube I9, provided with a special valve 20.- The rubber`Aconnection I8 is used so that the tube I9 and other portions of theapparatus attached thereto may be readily sealed oil. from the rest ofthe attachment or apparatus by a vacuum-tight seal. Valve 20, which isin- .cluded in tube I9, consists of a rotatable metal disc having anopening 2| adapted to receive a rubber stopper suitable for seal forsealing the container or vial from the upper portion of tube I9, and toturn it around and allow it to drop into tube I8. This Valve may beprovided with a small air hole 22 to prevent the rubber stopper fromadhering to the valve when itis turned. Connected to the tube I9 bymeans of rubber tube 23 is a tube 24, of glass or other material,adapted to hold a number of thel rubber stoppers, each upside down. Thetube is provided at the top with a stopper 25 having a smallair hole 26and an air filter made of absorbent cotton or similar material 21. Thuswhen a stopper is removed from this tube by means of the valve 28, thisair filter and air hole allow sufcient sterile air to` enter the tube toprevent the formation of a vacuum thereinwhich might interfere withfeeding the stoppers through the valve. This tube for the Stoppers issupported by means of a ring or simllar device 28, which is of aconstruction which will permit the tube to be taken olf or replaced atwill, and which will permit the tube to move vertically with the lowerportion ofthe attachment and with respect to the upper portion.

An advantageous method of fastening the attachments to a vacuummanifold, and a suitable manifold, are illustrated in Figs. 9 and 10. In

attachment is provided with a tube 24 for the rubber stoppers, and witha somewhat modified form of air filter 4I on this tube. This air filterincludes a glass tube 42, containing absorbent cotton or the like 43,attached to tube 24 by rubber connection 44. The extensible tube I3 isVfastened to a metal tube I2, as in Fig. .1, and this tube is providedwith a threaded stud or bolt 45 carrying a wing nut 46. A rubber tube 41is fastened to the other end of tube I2with a vacuumtight connection.

This rubber tube fits over an outlet 48 on vacuum manifold 49, which maybe provided with IIII) or more such outlets (Figs. 9 and 10). Eachoutlet 48 is provided with a. forked support 58, the slot of which fitsover the bolt or stud 45, the attachment then being secured in place bythe wing nut 46. In this way, the attachment is rigidly supported bytheV vacuum manifold, and, by means of tube 41, is connected thereto bya vacuum-tight connection. The outlets on the vacuum manifold projectupwardly at an angle,

so that the gases and vapors, which pass upwardly through the tube I3,and then pass downwardly through the tube 41 into the outlet 48 and intothe vacuum manifold, thus preventingthe contamination of the material inthe container by the contents of the manifold or its outlet.

In operation, the entire attachment is suitably wrapped and sterilized,a clamp being placed on the` rubber tube at the top of the attachment,and on t'he tube between the stopper receptacle and the tube 4. 'Ijhewrapping around the rubber tube 4at the top ofthe apparatus is removed,and the attachment fastened to the vacuum` manifold, suitableprecautions being taken to insure sterility. The rest of the wrapping isthen removed, and', if a bell is provided at the lower end of theattachment, as in Fig. 9, the open end of the bell is immersed inantiseptic solution. V

A container I having suitably frozen material Ia within it'is attachedas shown in Figs. 1 and 9, with sterile technique, the clamp' on theconnection to the vacuum manifold is removed and the vacuum is appliedand the material desiccated or dehydrated. After the desiccation ordehydration is complete, the clamp is removed from Ythe tube I 8, astopper 29 isintroduced into tube I8 by means of the valve, and theclamp is replaced on the tube I8. The lower portion of the apparatus,including the container, is then pulled down to the position shown inFig. 2, the extensible or iiexible tube I3 permitting this. When thelower portion of the apparatus is pulled down, the tube II, beingrigidly connected to tube I2, no longer blocks the opening betweensleeves 4 and I8, and permits the rubber stopper 29 to fall intoposition in the neck of the vial or container. As soon as the rubberstopper is in the proper position in the neck of the container, thelowerportion of the attachment or apparatus is raised, as shown in: Fig. 3,so that the plunger II extends down into the neck of vial or containerand forces the rubber stopper into tight engagement with the taperedportion of the neck of the vial, thus insuring. a. tight seal which willretain the high vacuum in the vial for such short periods of time as arerequired during the sealing of the glass neck of the vial' and duringthe restoration of the material to a liquid state.

After the rubber stopper has been forced into tight engagement with theneck of the vial while maintaining the vacuum therein, the rubber tubeconnecting the attachment to the vacuum manifold lsclamped, and theclamp on the rubber tube I-8, connecting the tube 24 and the tube 4 isremoved, permitting air to enter theattachment through the air filter.AIn this way, the vacuum in the attachment is broken without the insideof the, attachment being contaminated. The sealed vial, as in Fig. 4, isthen removed, and the bell again immersed in antiseptic solution. ThisVseries of operations may then be repeated several times, and' a numberof containers may be so processed, with .sterile technique, and withoutrequiring that the` attachment be sterilized each time a container isprocessed.

toration is desired. The neck of the vial or container may be sealedbefore the container is removed from the apparatus, but preferably the4sealing is accomplished afterward, as this simplifles 'the production ofa proper seal, and avoids 'the diiliculty encountered in sealing arelatively vacuum in the sealed neck between the rubber stopper and theglass seal, as this-is a particularly advantageous method of sealing thecontainers, and is the method which I prefer to use. The container soproduced, having a vacuum both in the container proper, and a` vacuum inthe neck between the rubber stopper and the glass seal is the form ofcontainer which I prefer and which is particularly advantageous.Nevertheless., my invention is not limited tol the production of suchcontainers, or Isuch containers,' as the glass seal may be made withoutconnecting the neck to a vacuum, and without producing a vacuum withinthe neck, particularly with those containers in which the volume of thecontainer is relatively large, and is many times as great as the volumeof-the neck, or that portion of the neck between the rubber stopper andthe glass seal, without departing from the scope of my invention. Whenthe neck is name-sealed without first producing a vacuum within theneck, the air Within is greatly attenuated bythe heat required Ato sealthe glass and isffree from any appreciable amount of moisture so thatthere is butlittle air present above the rubber stopper to penetratepast the stopper into the'container, and almost no moisture, so that,despite the fact that some air may enter the container, the amount whichcan enter is almost innitesimal, so that the vacuum within ,thecontainer, particularly where the container is vthe needle 32 of a.hypodermic syringe through the rubber stopper as shown in Figs. 7 and 8,forcing water through the needle into the evacu.

ated container. The rubber Stoppers are advanta'geously provided with apassage 33 (Fig. 3) extending part way through them to facilitate thepassage of the needle. After the proper amount of water hasbeen forcedinto the container and the material wi-thin the container is dissolved,the needle is removed temporarily to admit air into the container sothat the dissolved material can be removed, the needle of the hypodermicsyringe is again inserted through the rubber stopper and the containerand syringe turned upside down, as shown in'Fig. 8 and the liquidmaterial sucked into the hypodermic syringe, whereupon it is ready forinjection.

During such times as no container is attached to' the apparatus, asuitable clamp is placed on tube Il to form a vacuum-tight connection toprevent breaking the vacuum Within vthe vacuum manifold. A clamp iscustomarily used on rubber tube I8 to' insure a vacuum-tight connectionto prevent the leakage of air through stopper 25 and past valve 20 intotubes I l) and 4. This clamp is taken off to permit Stoppers to passfrom the valve 20 down` into the tube I0 at such times as are desired,and during such times, the valve v2|) is sufficiently tight to preventthe ingress of much air, and if the attachment is used in conjunctionwith a manifold of suitable size, the amount of air which enters at thetime of introduction of a stopper into tne tube I0 is so small thatthere is almost no noticeable change in the pressure Within theapparatus. This clamp may also be removed after a rubber stopper hasbeen inserted in a container, and when it is desired to remove thecontainer from the attachment, to permit breaking the vacuum in thecontainer with sterile air suitably ltered. When this is done, careshouldbe taken that the rubber tube connecting'- the attachmenty to thevacuum manifold is clamped. However, the clamp should not be left off ofthe rubber tube I8 for any longer period of time than is necessary.

It `will be understood that the apparatus as described above is by wayof illustration, and that many modifications and embodiments of theinvention may occur to those skilled in the art which are within thescope of the present invention. For example, instead of providing a tubeor cylinder containing a number of Stoppers which may be used forthe'sealing of a number of containers, a suitable tube containing butone stopper, and hence not requiring a mechanism such as the valve 20 tointroduce the Stoppers into the tube I0 singly, may be used. Theprovision of tinuous operation of the apparatus with a minimum of laborand a minimum o f changing of connections, etc., and simplies theintroduction of a sterile stopper into each container. For example, atube equipped with a suitable air filter may be supplied with some 7 or8 or more Stoppers, the entire tube and Stoppers being sterilized in asuitable manner, as by steam sterilization or by baking, and connectedto the apparatus by tube 23 with sterile precautions. -With the airfilter 21 provided, the sterility of the Stoppers is assured, and anumber of containers equal to the number of Stoppers contained in thetube may be processed without the requirement of ,sterilizing a stopperand container therefor each time a container is processed.

In carrying out a desiccation process such as described, theperiodrequired to desiccate the material is fairly prolonged, and it isdesirable to conduct'the operation so that for each attachu ment onecontainer may be processed per day. In order to accomplish this, itis-necessary to have the actual processing time not more than 22hours,leaving some 2 hours for sealing the containers and replacing themwith new containers with frozen material in them. The process andapparatus of the present invention greatly speeds the Vial.

up the changing of containers and the sealingV ofthe containers. Inorder to seal a container according to the process of the presentinvention,

lt is merely necessary to introduce a stopper into the neck of thecontainer by allowing a stopper to pass from the valve 2U into the tubeI0, pulling down the lower part ofthe apparatus sothat the stopper fallsinto the proper position in the neck of the vial, and then to push upthe lower part of the apparatus so that the plunger l I forces thestopper into tight engagement with the neck of The container may then beremoved from the apparatus, and a new container placed thereon, readyfor processing. It is not necessary to flame-seal the neck of thecontainer immediately, as during a short period of exposure the sealobtained by means of the rubber stopper is suiiicient to hold the highvacuum within the container and to prevent ingress of moisture or anycontamination.

While I have described the invention with particular reference to itsapplication to the production and sealing of desiccated biologicallyactive substances in a final container, it is not limited thereto; as inits broader aspects it includes the new process and apparatus as appliedto the introduction of a perforable stopper into the neck of a containerwhile maintaining a vacuum therein, and to the production of anAevacuated container having an all-glass seal, and

having a perforable closure sealed within it; the stopperbeing`introduced and forced into place while a vacuum is maintainedwithin the container. Nevertheless, the invention is of particularadvantage for the production of desiccated biologically activesubstances in a sealed container provided with a, glass seal, and with aperforable stopper sealed within it, such thatthe Vglass seal aiordsabsolute assurance that the vacuum within the container is maintainedand that the ingress of moisture or contaminating substances isprevented, and yet which is adapted for the introduction of water orliquid through the perforable stopper, after the glass seal is y broken,without breaking the vacuum.

-container bys gradually applied pressure, and y flame sealing the neckof the container above the perforable stopper.

2. The method of sealing desiccated biologically'active substances .in afinal glasscontainer i which comprises introducing a perforable stopperinto the neck of the container whilemaintaining a vacuum within thecontainer, and sealing the neck ofthe container above the perforablestop- `per while maintaining a vacuum inthe neck above .the perforablestopper.

y3. The method of sealing desiccated biologically active substances in'a final glasscontainer which comprises introducing a perforable stopperinto the neckof the container while maintaining a vacuum within'thecontainer, forcing said stopper into tight engagement therewith, andsealing the neck of the container'above the perforable stopper whilemaintaining a vacuum in the neck above the perforable stopper.

4. Apparatus adapted for sealing substances -in a `final glass containercomprising an extensible tube, a movable plunger within said tube, meansfor fastening a container to one end of said tube, and means including albranch from said tube adapted to be sealed with a vacuum tight seal andnormally blocked by said plunger for' introducing a perforable stopperwithin said tube and within the neck of the container.

5. Apparatus adapted for sealing substances in an extensible portion, amovable hollow plunger within said tube, means for connecting acontainer to one end of Isaid tube, means for applying a vacuum to theotherfend of said tube, means, including a branch from said tube adaptedto be sealed with a vacuum tight seal and normally blocked by saidplunger, for introducing a stopper into said tube and into the neck ofthe container, whereby it may 'be forced into tight engagement with theneck of the container by the plunger while a vacuum is maintained insaid container.

6. Apparatus adapted for sealing substances in a'iinal glass containercomprising a tube having an extensible portion, a hollow plunger, iixedwith respect to'one end of said tube and movk able with respect to theother, within saidtube,

engagement with the neck of the container by.

reversing the movement of the 4movable portion of the tube and causingsaid plunger to enter said ,neck and press against-the 'stopper while avacuum is maintained in said container.

7. A device for introducing stoppers into a tube comprising a receptacleadapted to receive a plu-r a final glass container comprising a tubehavinglao rality of stoppers, and a valve connected thereto I adapted toreceive a single stopper, reverse it, and deliver it to said tube.

8. A device for introducing stoppers into a tube comprising a receptacleadapted to receive a plurality of stoppers, and a valve connectedthereto adaptedto receive a single stopper, reverse it, and deliver itto said tube, and means for introducing sterile air into saidreceptacle.

9. A device for introducing stoppers into an evacuated tube comprising areceptacle adapted to receive a plurality of stoppers, a valve attachedthereto adapted to receive a single stopper, reverse it, and Vdeliver itto Vsaid tube, and means for making la vacuumtight seal between saidvalve and said tube.

10. The method of sealing a glass container which comprises introducinga perforable stopper into the-neck of the container while maintaining avacuum within the container, forcing said stopper into tight engagementwith the neck of the container to form a Avacuum tight seal, thenbreaking the vacuum in the neck of the container above the stopper,drawing a portion ofthe neck above the stopper to a relatively ne tube,re-

evacuating the neck and flame-sealing it at the drawn Iportion whilemaintaining a, vacuum therein, whereby both the container and the neckof the container above the stopper are maintained in an evacuatedcondition.

11. 'I'he method of sealing desiccated biologically active 'substancesin a nal glass container which comprises introducing a perforable stopiper-into the neck of the container while maintaining a vacuum within thecontainer, and flame sealing: the neck o f 'the container above theperforable stopper. 1

COURTLAND H. BARR.

